Metalloproteinase inhibitors

ABSTRACT

Compounds of the formula (I) wherein Z is SO 2 (N6) or N(R7)SO 2  or N(R7)SO 2 N(R6) 
                         
useful as metalloproteinase inhibitors, especially as inhibitors of MMP12.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of United States (“U.S.”) applicationSer. No. 12/114,901, filed on May 5, 2008 now U.S. Pat. No. 7,666,892,which is a divisional of U.S. application Ser. No. 10/471,810, filed onSep. 12, 2003 now U.S. Pat. No. 7,368,465, which is the US NationalStage under 35 U.S.C. §371 of International Application No.PCT/SE2002/00478, filed Mar. 13, 2002, which in turn claims the benefitof Swedish Application Serial No. 0100902-6, filed on Mar. 15, 2001.Each of each of these prior applications is incorporated herein byreference in its entirety.

The present invention relates to compounds useful in the inhibition ofmetalloproteinases and in particular to pharmaceutical compositionscomprising these, as well as their use.

The compounds of this invention are inhibitors of one or moremetalloproteinase enzymes. Metalloproteinases are a superfamily ofproteinases (enzymes) whose numbers in recent years have increaseddramatically. Based on structural and functional considerations theseenzymes have been classified into families and subfamilies as describedin N. M. Hooper (1994) FEBS Letters 354:1-6. Examples ofmetalloproteinases include the matrix metalloproteinases (MMPs) such asthe collagenases (MMP1, MMP8, MMP13), the gelatinases (MMP2, MMP9), thestromelysins (MMP3, MMP10, MMP11), matrilysin (MMP7), metalloelastase(MMP12), enamelysin (MMP19), the MT-MMPs (MMP14, MMP15, MMP16, MMP17);the reprolysin or adamalysin or MDC family which is includes thesecretases and sheddases such as TNF converting enzymes (ADAM10 andTACE); the astacin family which include enzymes such as procollagenprocessing proteinase (PCP); and other metalloproteinases such asaggrecanase, the endothelin converting enzyme family and the angiotensinconverting enzyme family.

Metalloproteinases are believed to be important in a plethora ofphysiological disease processes that involve tissue remodelling such asembryonic development, bone formation and uterine remodelling duringmenstruation. This is based on the ability of the metalloproteinases tocleave a broad range of matrix substrates such as collagen, proteoglycanand fibronectin. Metalloproteinases are also believed to be important inthe processing, or secretion, of biological important cell mediators,such as tumour necrosis factor (TNF); and the post translationalproteolysis processing, or shedding, of biologically important membraneproteins, such as the low affinity IgE receptor CD23 (for a morecomplete list see N. M. Hooper et al., (1997) Biochem J. 321:265-279).

Metalloproteinases have been associated with many diseases orconditions. Inhibition of the activity of one or more metalloproteinasesmay well be of benefit in these diseases or conditions, for example:various inflammatory and allergic diseases such as, inflammation of thejoint (especially rheumatoid arthritis, osteoarthritis and gout),inflammation of the gastro-intestinal tract (especially inflammatorybowel disease, ulcerative colitis and gastritis), inflammation of theskin (especially psoriasis, eczema, dermatitis); in tumour metastasis orinvasion; in disease associated with uncontrolled degradation of theextracellular matrix such as osteoarthritis; in bone resorptive disease(such as osteoporosis and Paget's disease); in diseases associated withaberrant angiogenesis; the enhanced collagen remodelling associated withdiabetes, periodontal disease (such as gingivitis), corneal ulceration,ulceration of the skin, post-operative conditions (such as colonicanastomosis) and dermal wound healing; demyelinating diseases of thecentral and peripheral nervous systems (such as multiple sclerosis);Alzheimer's disease; extracellular matrix remodelling observed incardiovascular diseases such as restenosis and atherosclerosis; asthma;rhinitis; and chronic obstructive pulmonary diseases (COPD).

MMP12, also known as macrophage elastase or metalloelastase, wasinitially cloned in the mouse by Shapiro et al [1992, Journal ofBiological Chemistry 267: 4664] and in man by the same group in 1995.MMP-12 is preferentially expressed in activated macrophages, and hasbeen shown to be secreted from alveolar macrophages from smokers[Shapiro et al, 1993, Journal of Biological Chemistry, 268: 23824] aswell as in foam cells in atherosclerotic lesions [Matsumoto et al, 1998,Am J Pathol 153: 109]. A mouse model of COPD is based on challenge ofmice with cigarette smoke for six months, two cigarettes a day six daysa week. Wildtype mice developed pulmonary emphysema after thistreatment. When MMP12 knock-out mice were tested in this model theydeveloped no significant emphysema, strongly indicating that MMP-12 is akey enzyme in the COPD pathogenesis. The role of MMPs such as MMP12 inCOPD (emphysema and bronchitis) is discussed in Anderson and Shinagawa,1999, Current Opinion in Anti-inflammatory and ImmunomodulatoryInvestigational Drugs 1(1): 29-38. It was recently discovered thatsmoking increases macrophage infiltration and macrophage-derived MMP-12expression in human carotid artery plaques Kangavari [Matetzky S,Fishbein M C et al., Circulation 102:(18), 36-39 Suppl. S, Oct. 31,2000].

MMP13, or collagenase 3, was initially cloned from a cDNA libraryderived from a breast tumour [J. M. P. Freije et al. (1994) Journal ofBiological Chemistry 269(24):16766-16773]. PCR-RNA analysis of RNAs froma wide range of tissues indicated that MMP13 expression was limited tobreast carcinomas as it was not found in breast fibroadenomas, normal orresting mammary gland, placenta, liver, ovary, uterus, prostate orparotid gland or in breast cancer cell lines (T47-D, MCF-7 and ZR75-1).Subsequent to this observation MMP13 has been detected in transformedepidermal keratinocytes [N. Johansson et al., (1997) Cell Growth Differ.8(2):243-250], squamous cell carcinomas [N. Johansson et al., (1997) Am.J. Pathol. 151(2):499-508] and epidermal tumours [K. Airola et al.,(1997) J. Invest. Dermatol. 109(2):225-231]. These results aresuggestive that MMP13 is secreted by transformed epithelial cells andmay be involved in the extracellular matrix degradation and cell-matrixinteraction associated with metastasis especially as observed ininvasive breast cancer lesions and in malignant epithelia growth in skincarcinogenesis.

Recent published data implies that MMP13 plays a role in the turnover ofother connective tissues. For instance, consistent with MMP13'ssubstrate specificity and preference for degrading type II collagen [P.G. Mitchell et al., (1996) J. Clin. Invest. 97(3):761-768; V. Knauper etal., (1996) The Biochemical Journal 271:1544-1550], MMP13 has beenhypothesised to serve a role during primary ossification and skeletalremodelling [M. Stahle-Backdahl et al., (1997) Lab. Invest.76(5):717-728; N. Johansson et al., (1997) Dev. Dyn. 208(3):387-397], indestructive joint diseases such as rheumatoid and osteo-arthritis [D.Wernicke et al., (1996) J. Rheumatol. 23:590-595; P. G. Mitchell et al.,(1996) J. Clin. Invest. 97(3):761-768; O. Lindy et al., (1997) ArthritisRheum 40(8):1391-1399]; and during the aseptic loosening of hipreplacements [S. Imai et al., (1998) J. Bone Joint Surg. Br.80(4):701-710]. MMP13 has also been implicated in chronic adultperiodontitis as it has been localised to the epithelium of chronicallyinflamed mucosa human gingival tissue [V. J. Uitto et al., (1998) Am. J.Pathol 152(6):1489-1499] and in remodelling of the collagenous matrix inchronic wounds [M. Vaalamo et al., (1997) J. Invest. Dermatol.109(1):96-101].

MMP9 (Gelatinase B; 92 kDa TypeIV Collagenase; 92 kDa Gelatinase) is asecreted protein which was first purified, then cloned and sequenced, in1989 [S. M. Wilhelm et al (1989) J. Biol Chem. 264 (29): 17213-17221;published erratum in J. Biol Chem. (1990) 265 (36): 22570]. A recentreview of MMP9 provides an excellent source for detailed information andreferences on this protease: T. H. Vu & Z. Werb (1998) (In: MatrixMetalloproteinases. 1998. Edited by W. C. Parks & R. P. Mecham. pp115-148. Academic Press. ISBN 0-12-545090-7). The following points aredrawn from that review to by T. H. Vu & Z. Werb (1998).

The expression of MMP9 is restricted normally to a few cell types,including trophoblasts, osteoclasts, neutrophils and macrophages.However, it's expression can be induced in these same cells and in othercell types by several mediators, including exposure of the cells togrowth factors or cytokines. These are the same mediators oftenimplicated in initiating an inflammatory response. As with othersecreted MMPs, MMP9 is released as an inactive Pro-enzyme which issubsequently cleaved to form the enzymatically active enzyme. Theproteases required for this activation in vivo are not known. Thebalance of active MMP9 versus inactive enzyme is further regulated invivo by interaction with TIMP-1 (Tissue Inhibitor ofMetalloproteinases-1), a naturally-occurring protein. TIMP-1 binds tothe C-terminal region of MMP9, leading to inhibition of the catalyticdomain of MMP9. The balance of induced expression of ProMMP9, cleavageof Pro- to active MMP9 and the presence of TIMP-1 combine to determinethe amount of catalytically active MMP9 which is present at a localsite. Proteolytically active MMP9 attacks substrates which includegelatin, elastin, and native Type IV and Type V collagens; it has noactivity against native Type I collagen, proteoglycans or laminins.

There has been a growing body of data implicating roles for MMP9 invarious physiological and pathological processes. Physiological rolesinclude the invasion of embryonic trophoblasts through the uterineepithelium in the early stages of embryonic implantation; some role inthe growth and development of bones; and migration of inflammatory cellsfrom the vasculature into tissues.

MMP-9 release, measured using enzyme immunoassay, was significantlyenhanced in fluids and in AM supernantants from untreated asthmaticscompared with those from other populations [Am. J. Resp. Cell & Mol.Biol., November 1997, 17 (51:583-591]. Also, increased MMP9 expressionhas been observed in certain other pathological conditions, therebyimplicating MMP9 in disease processes such as COPD, arthritis, tumourmetastasis, Alzheimer's, Multiple Sclerosis, and plaque rupture inatherosclerosis leading to acute coronary conditions such as MyocardialInfarction.

MMP-8 (collagenase-2, neutrophil collagenase) is a 53 kD enzyme of thematrix metalloproteinase family that is preferentially expressed inneutrophils. Later studies indicate MMP-8 is expressed also in othercells, such as osteoarthritic chondrocytes [Shlopov et al, 1997,Arthritis Rheum, 40:2065]. MMPs produced by neutrophils can cause tissueremodelling, and hence blocking MMP-8 should have a positive effect infibrotic diseases of for instance the lung, and in degradative diseaseslike pulmonary emphysema. MMP-8 was also found to be up-regulated inosteoarthritis, indicating that blocking MMP-8 many also be beneficialin this disease.

MMP-3 (stromelysin-1) is a 53 kD enzyme of the matrix metalloproteinaseenzyme family. MMP-3 activity has been demonstrated in fibroblastsisolated from inflamed gingiva [Uitto V. J. et al, 1981, J. PeriodontalRes., 16:417-424], and enzyme levels have been correlated to theseverity of gum disease [Overall C. M. et al, 1987, J. Periodontal Res.,22:81-88]. MMP-3 is also produced by basal keratinocytes in a variety ofchronic ulcers [Saarialho-Kere U. K. et al, 1994, J. Clin. Invest.,94:79-88]. MMP-3 mRNA and protein were detected in basal keratinocytesadjacent to but distal from the wound edge in what probably representsthe sites of proliferating epidermis. MMP-3 may thus prevent theepidermis from healing. Several investigators have demonstratedconsistent elevation of MMP-3 in synovial fluids from rheumatoid andosteoarthritis patients as compared to controls [Walakovits L. A. et al,1992, Arthritis Rheum., 35:35-42; Zafarullah M. et al, 1993, J.Rheumatol., 20:693-697]. These studies provided the basis for the beliefthat an inhibitor of MMP-3 will treat diseases involving disruption ofextracellular matrix resulting in inflammation due to lymphocyticinfiltration, or loss of structural integrity necessary for organfunction.

A number of metalloproteinase inhibitors are known (see for example thereview of MMP inhibitors by Beckett R. P. and Whittaker M., 1998, Exp.Opin. Ther. Patents, 8(3):259-282]. Different classes of compounds mayhave different degrees of potency and selectivity for inhibiting variousmetalloproteinases.

Whittaker M. et al (1999, Chemical Reviews 99(9):2735-2776] review awide range of known MMP inhibitor compounds. They state that aneffective MMP inhibitor requires a zinc binding group or ZBG (functionalgroup capable of chelating the active site zinc(II) ion), at least onefunctional group which provides a hydrogen bond interaction with theenzyme backbone, and one or more side chains which undergo effective vander Waals interactions with the enzyme subsites. Zinc binding groups inknown MMP inhibitors include carboxylic acid groups, hydroxamic acidgroups, sulfhydryl or mercapto, etc. For example, Whittaker M. et aldiscuss the following MMP inhibitors:

The above compound entered clinical development. It has a mercaptoacylzinc binding group, a trimethylhydantoinylethyl group at the P1 positionand a leucinyl-tert-butyllglycinyl backbone.

The above compound has a mercaptoacyl zinc binding group and an imidegroup at the P1 position.

The above compound was developed for the treatment of arthritis. It hasa non-peptidic succinyl hydroxamate zinc binding group and atrimethylhydantoinylethyl group at the P1 position.

The above compound is a phthalimido derivative that inhibitscollagenases. It has a non-peptidic succinyl hydroxamate zinc bindinggroup and a cyclic imide group at P1. Whittaker M. et al also discussother MMP inhibitors having a P1 cyclic imido group and various zincbinding groups (succinyl hydroxamate, carboxylic acid, thiol group,phosphorous-based group).

The above compounds appear to be good inhibitors of MMP8 and MMP9 (PCTpatent applications WO9858925, WO9858915). They have apyrimidin-2,3,4-trione zinc binding group.

The following compounds are not known as MMP inhibitors:—

Lora-Tamayo, M et al (1968, An. Quim 64(0: 591-606) describe synthesisof the following compounds as a potential anti-cancer agent:

Czech patent numbers 151744 (19731119) and 152617 (1974022) describe thesynthesis and the anticonvulsive activity of the following compounds:

U.S. Pat. No. 3,529,019 (19700915) describes the following compoundsused as intermediates:

PCT patent application number WO 00/09103 describes compounds useful fortreating a vision disorder, including the following (compounds 81 and83, Table A, page 47):

We have now discovered a new class of compounds that are inhibitors ofmetalloproteinases and are of particular interest in inhibiting MMPssuch as MMP-12. The compounds are metalloproteinase inhibitors having ametal binding group that is not found in known metalloproteinaseinhibitors. In particular, we have discovered compounds that are potentMMP12 inhibitors and have desirable activity profiles. The compounds ofthis invention have beneficial potency, selectivity and/orpharmacokinetic properties.

The metalloproteinase inhibitor compounds of the invention comprise ametal binding group and one or more other functional groups or sidechains characterised in that the metal binding group has the formula (k)

wherein

X is selected from NR1, O, S;

Y1 and Y2 are independently selected from O, S;

R1 is selected from H, alkyl, haloalkyl;

Any alkyl groups outlined above may be straight chain or branched; anyalkyl group outlined above is preferably (C1-7)alkyl and most preferably(C1-6)alkyl.

A metalloproteinase inhibitor compound is a compound that inhibits theactivity of a metalloproteinase enzyme (for example, an MMP). By way ofnon-limiting example the inhibitor compound may show IC50s in vitro inthe range of 0.1-10000 nanomolar, preferably 0.1-1000 nanomolar.

A metal binding group is a functional group capable of binding the metalion within the active site of the enzyme. For example, the metal bindinggroup will be a zinc binding group in MMP inhibitors, binding the activesite zinc(II) ion. The metal binding group of formula (k) is based on afive-membered ring structure and is preferably a hydantoin group, mostpreferably a −5 substituted 1-H,3-H-imidazolidine-2,4-dione.

In a first aspect of the invention we now provide compounds of theformula I

wherein

X is selected from NR1, O, S;

Y1 and Y2 are independently selected from O, S;

Z is selected from SO₂N(R6), N(R7)SO₂, N(R7)SO₂N(R6);

m is 1 or 2;

A is selected from a direct bond, (C1-6)alkyl, (C1-6)haloalkyl, or(C1-6)heteroalkyl to containing a hetero group selected from N, O, S,SO, SO2 or containing two hetero groups selected from N, O, S, SO, SO2and separated by at least two carbon atoms;

R1 is selected from H, (C1-3)alkyl, haloalkyl;

Each R2 and R3 is independently selected from H, halogen (preferablyfluorine), alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl,heteroaryl, alkylaryl, alkyl-heteroaryl, heteroalkyl-aryl,heteroalkyl-heteroaryl, aryl-alkyl, aryl-heteroalkyl, heteroaryl-alkyl,heteroaryl-heteroalkyl, aryl-aryl, aryl-heteroaryl, heteroaryl-aryl,heteroaryl-heteroaryl, cycloalkyl-alkyl, heterocycloalkyl-alkyl;

Each R4 is independently selected from H, halogen (preferably fluorine),(C1-3)alkyl or haloalkyl;

R6 is selected from H, alkyl, heteroalkyl, heterocycloalkyl, aryl,heteroaryl, alkylaryl, alkyl-heteroaryl, heteroalkyl-aryl,heteroalkyl-heteroaryl, arylalkyl, aryl-heteroalkyl, heteroaryl-alkyl,heteroaryl-heteroalkyl, aryl-aryl, aryl-heteroaryl, heteroaryl-aryl,heteroaryl-heteroaryl;

Each of the R2, R3 and R6 radicals may be independently optionallysubstituted with one or more (preferably one) groups selected fromalkyl, heteroalkyl, aryl, heteroaryl, halo, haloalkyl, hydroxy, alkoxy,haloalkoxy, thiol, alkylthiol, arylthiol, alkylsulfon, haloalkylsulfon,arylsulfon, aminosulfon, N-alkylaminosulfon, N,N-dialkylaminosulfon,arylaminosulfon, amino, N-alkylamino, N,N-dialkylamino, amido,N-alkylamido, N,N-dialkylamido, cyano, sulfonamino, alkylsulfonamino,arylsulfonamino, amidino, N-aminosulfon-amidino, guanidino,N-cyano-guanidino, thioguanidino, 2-nitro-ethene-1,1-diamin, carboxy,alkyl-carboxy, nitro;

Optionally R2 and R3 may join to form a ring comprising up to 7 ringatoms, or R2 and R4 may join to form a ring comprising up to 7 ringatoms, or R2 and R6 may join to form a ring comprising up to 7 ringatoms, or R3 and R4 may join to form a ring comprising up to 7 ringatoms, or R3 and R6 may join to form a ring comprising up to 7 ringatoms, or R4 and R6 may join to form a ring comprising up to 7 ringatoms;

R5 is a monocyclic, bicyclic or tricyclic group comprising one, two orthree ring structures each of up to 7 ring atoms independently selectedfrom cycloalkyl, aryl, heterocycloalkyl or heteroaryl, with each ringstructure being independently optionally substituted by one or moresubstituents independently selected from halogen, hydroxy, alkyl,alkoxy, haloalkoxy, amino, N-alkylamino, N,N-dialkylamino,alkylsulfonamino, alkylcarboxyamino, cyano, nitro, thiol, alkylthiol,alkylsulfonyl, haloalkylsulfonyl, alkylaminosulfonyl, carboxylate,alkylcarboxylate, aminocarboxy, N-alkylamino-carboxy,N,N-dialkylamino-carboxy, wherein any alkyl radical within anysubstituent may itself be optionally substituted with one or more groupsselected from halogen, hydroxy, alkoxy, haloalkoxy, amino, N-alkylamino,N,N-dialkylamino, N-alkylsulfonamino, N-alkylcarboxyamino, cyano, nitro,thiol, alkylthiol, alkylsulfonyl, N-alkylaminosulfonyl, carboxylate,alkylcarboxy, aminocarboxy, N-alkylaminocarboxy,N,N-dialkylaminocarboxy;

when R5 is a bicyclic or tricyclic group, each ring structure is joinedto the next ring structure by a direct bond, by —O—, by (C1-6)alkyl, by(C1-6)haloalkyl, by (C1-6)heteroalkyl, by (C1-6)alkenyl, by(C1-6)alkynyl, by sulfone, or is fused to the next ring structure;

R7 is selected from (C1-6) alkyl, (C3-7)cycloalkyl, (C2-6)heteroalkyl,(C2-6)cycloheteroalkyl;

Any heteroalkyl group outlined above is a hetero atom-substituted alkylcontaining one or more hetero groups independently selected from N, O,S, SO, SO2, (a hetero group being a hetero atom or group of atoms);

Any heterocycloalkyl or heteroaryl group outlined above contains one ormore hetero groups independently selected from N, O, S, SO, SO2;

Any alkyl, alkenyl or alkynyl groups outlined above may be straightchain or branched; unless otherwise stated, any alkyl group outlinedabove is preferably (C1-7)alkyl and most preferably (C1-6)alkyl;

Provided that:

-   -   when X is NR1, R1 is H, Y1 is O, Y2 is O, Z is SO₂N(R6), R6 is        H, R2 is H, m is 1, R3 is H, R4 is H, and A is a direct bond,        then R5 is not phenyl, p-nitro-phenyl, p-ethoxyphenyl or        m-methylphenyl;    -   when X is S or NR1 and R1 is H, Y1 is O, Y2 is O, Z is SO₂N(R6),        R6 is alkyl, R2 is H, m is 1, one of R3 and R4 is H and the        other is alkyl, R3 and R6 or R4 and R6 join to form a 5-membered        ring, and A is a direct bond, then R5 is not phenyl.

Preferred compounds of the formula I are those wherein any one or moreof the following apply:

-   -   X is NR1;    -   Z is SO₂N(R6), especially wherein the S atom of group Z is        attached to group A in the compound of formula I;    -   At least one of Y1 and Y2 is O; especially both Y1 and Y2 are O;    -   m is 1;    -   R1 is H, (C1-3) alkyl, (C1-3) haloalkyl; especially R1 is H;

R2 is H, alkyl, hydroxyalkyl, aminoalkyl, cycloalkyl-alkyl,alkyl-cycloalkyl, arylalkyl, alkylaryl, heteroalkyl,heterocycloalkyl-alkyl, alkyl-heterocycloalkyl, heteroaryl-alkyl,heteroalkyl-aryl; especially R2 is alkyl, aminoalkyl orheteroaryl-alkyl.

R3 and/or R4 is H;

R3 and/or R4 is methyl;

R3 and R4 form a 5- or 6-membered ring (preferably a 5-membered ring) orR3 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring) orR4 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring);especially R3 and R6 form a 5- or 6-membered ring, most preferably a5-membered ring;

R2 and R3 form a 5-membered ring or R2 and R6 form a 5-membered ring;

R5 comprises one, two or three optionally substituted aryl or heteroaryl5- or 6-membered rings;

R5 is a bicyclic or tricyclic group comprising two or three optionallysubstituted ring structures;

R3 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring) orR4 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring)and R5 is a bicyclic or tricyclic group comprising two or threeoptionally substituted ring structures.

Particularly preferred compounds of formula I are those wherein R5 is abicyclic or tricyclic group comprising two or three optionallysubstituted ring structures.

For example, particular compounds of formula I are those wherein Y1 isO, Y2 is O, X is NR1, R1 is H, R2 is H, m is 1, R3 is H, R4 is H, Z isSO₂N(R6), R6 is H, (C1-4)alkyl, methylbenzyl, or methylpyridyl, A is adirect bond, and R5 is a bicyclic or tricyclic group comprising two orthree optionally substituted ring structures. Some such compounds aredescribed in Examples 1 and 2.

Other particular compounds of formula I are those wherein Y1 is O, Y2 isO, X is NR1, R1 is H, R2 is H, methyl, or benzyl, m is 1, R3 is H ormethyl, R4 is H, Z is SO₂N(R6), R6 is H, A is a direct bond, and R5 is abicyclic or tricyclic group comprising two or three optionallysubstituted ring structures. Some such compounds are described inExample 3.

The invention further provides compounds of the formula II

wherein

each of G1 and G2 is a monocyclic ring structure comprising each of upto 7 ring atoms independently selected from cycloalkyl, aryl,heterocycloalkyl or heteroaryl, with each ring structure beingindependently optionally substituted by one or two substituentsindependently selected from halogen, hydroxy, haloalkoxy, amino,N-alkylamino, N,N-dialkylamino, cyano, nitro, alkyl, alkoxy, alkylsulfone, haloalkyl sulfone, alkylcarbamate, alkylamide, wherein anyalkyl radical within any substituent may itself be optionallysubstituted with one or more groups selected from halogen, hydroxy,amino, N-alkylamino, N,N-dialkylamino, cyano, nitro, alkoxy, haloalkoxy;

Z is SO₂N(R6);

B is selected from a direct bond, O, (C1-6)alkyl, (C1-6)heteroalkyl;

R2 is selected from H, (C1-6)alkyl, haloalkyl, hydroxyalkyl,alkoxyalkyl, aminoalkyl, (N-alkylamino)alkyl, (N,N-dialkylamino)alkyl,amidoalkyl, thioalkyl, or R2 is a group of formula III

C and D are independently selected from a direct bond, H, (C1-C6)alkyl,(C1-C6)haloalkyl, or (C1-C6)heteroalkyl containing one or two heteroatoms selected from N, O or S such that when two hetero atoms arepresent they are separated by at least two carbon atoms;

G3 is a monocyclic ring structure comprising up to 7 ring atomsindependently selected from cycloalkyl, aryl, heterocycloalkyl orheteroaryl, optionally substituted by one or two substituentsindependently selected from halogen, hydroxy, amino, N-alkylamino,N,N-dialkylamino, cyano, nitro, alkyl, alkoxy, alkyl sulfone, haloalkylsulfone, or alkyl substituted with one or more groups selected fromhalogen, hydroxy, amino, N-alkylamino, N,N-dialkylamino, cyano, nitro,alkoxy, haloalkoxy;

Optionally R2 is substituted with halo, haloalkyl, hydroxy, alkoxy,haloalkoxy, amino, aminoalkyl, N-alkylamino, N,N-dialkylamino,(N-alkylamino)alkyl, (N,N-dialkylamino)alkyl, alkylsulfone,aminosulfone, N-alkylamino-sulfone, N,N-dialkylamino-sulfone, amido,N-alkylamido, N,N-dialkylamido, cyano, sulfonamino, alkyl-sulfonamino,amidino, N-aminosulfone-amidino, guanidino, N-cyano-guanidino,thioguanidino, 2-nitroguanidino, 2-nitro-ethene-1,1-diamino, carboxy,alkylcarboxy;

R3 and R4 are independently selected from H or (C1-3)alkyl;

R6 is selected from H, (C1-3)alkylamino, or R6 is (C1-3)alkyl optionallysubstituted by aryl, heteroaryl, heterocycloalkyl;

Optionally R2 and R3 may join to form a ring comprising up to 7 ringatoms, or R2 and R4 may join to form a ring comprising up to 7 ringatoms, or R2 and R6 may join to form a ring comprising up to 7 ringatoms, or R3 and R4 may join to form a ring comprising up to 7 ringatoms, or R3 and R6 may join to form a ring comprising up to 7 ringatoms, or R4 and R6 may join to form a ring comprising up to 7 ringatoms;

Any heteroalkyl group outlined above is a hetero atom-substituted alkylcontaining one or more hetero groups independently selected from N, O,S, SO, SO2, (a hetero group being a hetero atom or group of atoms);

Any heterocycloalkyl or heteroaryl group outlined above contains one ormore hetero groups independently selected from N, O, S, SO, SO2;

Any alkyl, alkenyl or alkynyl groups outlined above may be straightchain or branched; unless otherwise stated, any alkyl group outlinedabove is preferably (C1-7)alkyl and most preferably (C1-6)alkyl.

Preferred compounds of the formula II are those wherein one or more ofthe following apply:

Z is SO₂N(R6) and the S atom of group Z is attached to the G2 ring;

B is a direct bond or O;

R2 is not optionally substituted, or R2 is selected from H, (C1-6)alkyl,aryl-(C1-6)alkyl or heteroaryl-(C1-6)alkyl optionally substituted withhalo, haloalkyl, hydroxy, alkoxy, haloalkoxy, amino, aminoalkyl,N-alkylamino, N,N-dialkylamino, (N-alkylamino)alkyl,(N,N-dialkylamino)alkyl, alkylsulfone, aminosulfone,N-alkylamino-sulfone, N,N-dialkylamino-sulfone, amido, N-alkylamido,N,N-dialkylamido, cyano, sulfonamino, alkyl-sulfonamino, amidino,N-aminosulfone-amidino, guanidino, N-cyano-guanidino, thioguanidino,2-nitroguanidino, 2-nitro-ethene-1,1-diamino, caboxy, alkylcarboxy;

Each of R3 and R4 is H;

R6 is H, benzyl or methylenepyridine;

G1 and G2 are each selected from an aryl or a heteroaryl;

R3 and R4 form a 5- or 6-membered ring (preferably a 5-membered ring) orR3 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring) orR4 and R6 form a 5- or 6-membered ring (preferably a 5-membered ring);especially R3 and R6 form a 5- or 6-membered ring, most preferably a5-membered ring;

R2 and R3 form a 5-membered ring or R2 and R6 form a 5-membered ring.

Particularly preferred compounds of the formula II are those wherein Zis SO₂N(R6) and the S atom of group Z is attached to the G2 ring.

For example, particular compounds of the invention include compounds offormula II wherein:

-   -   (a) B is a direct bond or O; and Z is SO2N(R6); and R2 is        selected from H, (C1-6)alkyl, aryl-(C1-6)alkyl or        heteroaryl-(C1-6)alkyl optionally substituted with halo,        haloalkyl, hydroxy, alkoxy, haloalkoxy, amino, aminoalkyl,        N-alkylamino, N,N-dialkylamino, (N-alkylamino)alkyl,        (N,N-dialkylamino)alkyl, alkylsulfonyl, aminosulfonyl,        N-alkylamino-sulfonyl, N,N-dialkylamino-sulfonyl, amido,        N-alkylamido, N,N-dialkylamido, cyano, sulfonamino,        alkyl-sulfonamino, amidino, N-aminosulfone-amidino, guanidino,        N-cyano-guanidino, thioguanidino, 2-nitroguanidino,        2-nitro-ethene-1,1-diamino, caboxy, alkylcarboxy; and each of R3        and R4 is H; and R6 is H, benzyl or methylenepyridine; or    -   (b) Z is SO₂N(R6), and R3 is H, and R4 is H (compounds of the        formula II′) wherein R2 is not optionally substituted;        preferably G1 and G2 are each selected from an aryl or a        heteroaryl:

Suitable values for R2 include the following:

Suitable values for R5 include the following:

It will be appreciated that the particular substituents and number ofsubstituents in compounds of the invention are selected so as to avoidsterically undesirable combinations.

Each exemplified compound represents a particular and independent aspectof the invention.

Where optically active centres exist in the compounds of the invention,we disclose all individual optically active forms and combinations ofthese as individual specific embodiments of the invention, as well astheir corresponding racemates. Racemates may be separated intoindividual optically active forms using known procedures (cf. AdvancedOrganic Chemistry: 3rd Edition: author J March, p 104-107) including forexample the formation of diastereomeric derivatives having convenientoptically active auxiliary species followed by separation and thencleavage of the auxiliary species.

It will be appreciated that the compounds according to the invention maycontain one or more asymmetrically substituted carbon atoms. Thepresence of one or more of these asymmetric centres (chiral centres) ina compound of formula I can give rise to stereoisomers, and in each casethe invention is to be understood to extend to all such stereoisomers,including enantiomers and diastereomers, and mixtures including racemicmixtures thereof.

Where tautomers exist in the compounds of the invention, we disclose allindividual tautomeric forms and combinations of these as individualspecific embodiments of the invention.

As previously outlined the compounds of the invention aremetalloproteinase inhibitors, in particular they are inhibitors ofMMP12. Each of the above indications for the compounds of the formula Irepresents an independent and particular embodiment of the invention.

Certain compounds of the invention are of particular use as inhibitorsof MMP13 and/or MMP9 and/or MMP8 and/or MMP3.

Compounds of the invention show a favourable selectivity profile. Whilstwe do not wish to be bound by theoretical considerations, the compoundsof the invention are believed to show selective inhibition for any oneof the above indications relative to any MMP1 inhibitory activity, byway of non-limiting example they may show 100-1000 fold selectivity overany MMP1 inhibitory activity.

The compounds of the invention may be provided as pharmaceuticallyacceptable salts. These include acid addition salts such ashydrochloride, hydrobromide, citrate and maleate salts and salts formedwith phosphoric and sulphuric acid. In another aspect suitable salts arebase salts such as an alkali metal salt for example sodium or potassium,an alkaline earth metal salt for example calcium or magnesium, ororganic amine salt for example triethylamine.

They may also be provided as in vivo hydrolysable esters. These arepharmaceutically acceptable esters that hydrolyse in the human body toproduce the parent compound. Such esters can be identified byadministering, for example intravenously to a test animal, the compoundunder test and subsequently examining the test animal's body fluids.Suitable in vivo hydrolysable esters for carboxy include methoxymethyland for hydroxy include formyl and acetyl, especially acetyl.

In order to use a metalloproteinase inhibitor compound of the invention(a compound of the formula I or II) or a pharmaceutically acceptablesalt or in vivo hydrolysable ester thereof for the therapeutic treatment(including prophylactic treatment) of mammals including humans, it isnormally formulated in accordance with standard pharmaceutical practiceas a pharmaceutical composition.

Therefore in another aspect the present invention provides apharmaceutical composition which comprises a compound of the invention(a compound of the formula I or II) or a pharmaceutically acceptablesalt or an in vivo hydrolysable ester and pharmaceutically acceptablecarrier.

The pharmaceutical compositions of this invention may be administered instandard manner for the disease or condition that it is desired totreat, for example by oral, topical, parenteral, buccal, nasal, vaginalor rectal administration or by inhalation. For these purposes thecompounds of this invention may be formulated by means known in the artinto the form of, for example, tablets, capsules, aqueous or oilysolutions, suspensions, emulsions, creams, ointments, gels, nasalsprays, suppositories, finely divided powders or aerosols forinhalation, and for parenteral use (including intravenous, intramuscularor infusion) sterile aqueous or oily solutions or suspensions or sterileemulsions.

In addition to the compounds of the present invention the pharmaceuticalcomposition of this invention may also contain, or be co-administered(simultaneously or sequentially) with, one or more pharmacologicalagents of value in treating one or more diseases or conditions referredto hereinabove.

The pharmaceutical compositions of this invention will normally beadministered to humans so that, for example, a daily dose of 0.5 to 75mg/kg body weight (and preferably of 0.5 to 30 mg/kg body weight) isreceived. This daily dose may be given in divided doses as necessary,the precise amount of the compound received and the route ofadministration depending on the weight, age and sex of the patient beingtreated and on the particular disease or condition being treatedaccording to principles known in the art.

Typically unit dosage forms will contain about 1 mg to 500 mg of acompound of this invention.

Therefore in a further aspect, we provide a compound of the formula I ora pharmaceutically acceptable salt or in vivo hydrolysable ester thereoffor use in a method of therapeutic treatment of the human or animal bodyor for use as a therapeutic agent. We disclose use in the treatment of adisease or condition mediated by one or more metalloproteinase enzymes.In particular we disclose use in the treatment of a disease or conditionmediated by MMP12 and/or MMP13 and/or MMP9 and/or MMP8 and/or to MMP3;especially use in the treatment of a disease or condition mediated byMMP12 or MMP9; most especially use in the treatment of a disease orcondition mediated by MMP12.

In particular we provide a compound of the formula II or apharmaceutically acceptable salt or in vivo hydrolysable ester thereoffor use in a method of therapeutic treatment of the human or animal bodyor for use as a therapeutic agent (such as use in the treatment of adisease or condition mediated by MMP12 and/or MMP13 and/or MMP9 and/orMMP8 and/or MMP3; especially MMP12 or MMP9; most especially MMP12).

In yet a further aspect we provide a method of treating ametalloproteinase mediated disease or condition which comprisesadministering to a warm-blooded animal a therapeutically effectiveamount of a compound of the formula I or a pharmaceutically acceptablesalt or in vivo hydrolysable ester thereof. We also disclose the use ofa compound of the formula I or a pharmaceutically acceptable salt or invivo hydrolysable precursor thereof in the preparation of a medicamentfor use in the treatment of a disease or condition mediated by one ormore metalloproteinase enzymes.

For example we provide a method of treating a metalloproteinase mediateddisease or condition which comprises administering to a warm-bloodedanimal a therapeutically effective amount of a compound of the formulaII (or a pharmaceutically acceptable salt or in vivo hydrolysable esterthereof). We also provide the use of a compound of the formula II (or apharmaceutically acceptable salt or in vivo hydrolysable precursorthereof) in the preparation of a medicament for use in the treatment ofa disease or condition mediated by one or more metalloproteinaseenzymes.

Metalloproteinase mediated diseases or conditions include asthma,rhinitis, chronic obstructive pulmonary diseases (COPD), arthritis (suchas rheumatoid arthritis and osteoarthritis), atherosclerosis andrestenosis, cancer, invasion and metastasis, diseases involving tissuedestruction, loosening of hip joint replacements, periodontal disease,fibrotic disease, infarction and heart disease, liver and renalfibrosis, endometriosis, diseases related to the weakening of theextracellular matrix, heart failure, aortic aneurysms, CNS relateddiseases such as Alzheimer's disease and Multiple Sclerosis (MS),hematological disorders.

Preparation of the Compounds of the Invention

In another aspect the present invention provides a process for preparinga compound of the formula I or II or a pharmaceutically acceptable saltor in vivo hydrolysable ester thereof, as described in (a) to (c) below.It will be appreciated that many of the relevant starting materials arecommercially or otherwise available or may be synthesised by knownmethods or may be found in the scientific literature.

(a) Compounds of formula I in which Y1 and Y2 are each 0, Z is SO2N(R6),A is a direct bond, X is NR1, R1 is H, R2 is H, m is 1, R3 is H, R4 isH, and R5 and R6 are defined as in formula I may be prepared accordingto Scheme 1.

When R6 is H, an N¹-BOC-D-diaminopropionic acid derivative of formula IVis reacted with suitable sulfonyl chloride of formula V in basic mediumto form sulfonamides of formula VI. Deprotection in acid medium,reaction with potassium cyanate to the corresponding urea and finallycyclization in acid medium yields compounds of formula I.

When R6 is alkyl such as methyl, ethyl, propyl, isopropyl and n-butyl,the N²-alkyl-N¹-BOC-D-diaminopropionic acid of formula IV is preparedaccording to Andruszkiewics, R.: Pol. J. Chem, 62, 257, (1988).

When R6 is an optionally substituted benzyl, methylbenzyl,methylpyridyl, methyl heteroaryl, the N²-substituted amino acid offormula IV is prepared according to Helv. Chim. Acta, 46, 327, (1963).

The reaction IV-VI is preferably performed in suitable solventoptionally in the presence of base for 1 to 24 h at ambient to refluxtemperature. Preferably, solvents such as pyridine, dimethylformamide,tetrahydrofurane, acetonitrile or dichlorometane are used with baseslike triethylamine, N-methylmorpholine, pyridine or alkali metalcarbonates at ambient temperature for 2-16 h reaction time, or until endof reaction is achieved as detected by chromatographic or spectroscopicmethods. Reactions of sulfonyl chlorides of formula V with varioussecondary amines are previously described in the literature, and thevariations of the conditions will be evident for those skilled in theart. A variety of compounds of formula V are commercially available ortheir synthesis is described in the literature. Specific derivatives offormula VI may be made according to known processes by those skilled inthe art.

(b) Compounds of formula I in which Y1 and Y2 are each 0, Z is SO2N(R6),R6 is H, A is a direct bond, X is NR1, R1 is H, m is 1, and R2, R3, R4and R5 are defined as in formula I may be prepared according to Scheme1.

Compounds in which R2 is H, R3 is H and R4 is alkyl or aryl, may beprepared starting from the corresponding BOC N-protected α-aminoaldehydes of formula VII, prepared according to Fehrentz, J A, Castro,B.; Synthesis, 676, (1983).

Compounds in which R2 is alkyl or aryl, R3 is H and R4 is alkyl or aryl,may be prepared starting from the corresponding BOC N-protected α-aminoketone of formula VII as depicted in Scheme 2. The BOC N-protectedα-amino ketones are prepared according to Nahm, S, Weinreb, S M:Tetrahedron Lett. 22, 3815, (1981), optionally when R6 is not H,according to Shuman, Robert T. U.S. Pat. No. 4,448,717 A 19840515

Some compounds prepared by the process shown in Scheme 2 are describedin Example 3.

The compounds of formula VII are reacted with alkali cyanide andammonium carbonate (Strecker reaction) to yield the correspondinghydantoins of formula VIIa. The diastereoisomeres can optionally beseparated after any of the three remaining synthetic steps: carbamatesof formula VIIa and sulfonamide compounds of formula I on silicagelchromatography, after deprotection amino intermediate bychrystallisation. The amine intermediates are optionally used todirectly couple with sulfonyl chlorides of formula V as described in thesulfonylation in (a) above, in basic medium to form compounds of formulaI.

The reaction VII to VIIa is preferably run in a closed steel vessel inan aqueous alcohol solvent at 90-130° C. for 3-16 hours or until end ofreaction is achieved as detected by chromatographic or spectroscopicmethods. Treatment with 1-4 fold excess cyanide salts, preferrably 1-2equivalents, and 2-6 fold excess of ammonium carbonate, preferrably 4-6equivalents yields hydantoins of formula VIIa. Deprotection andsulfonylation as in Scheme 1 then yields compounds of formula I.

Amino aldehydes or ketones of formula VII and their protectedderivatives are commercially available and other methods to α-aminoaldehydes and ketones of formula VII. Specific derivatives of formulaVIIa may be made according to known processes by those skilled in theart.

(c) Compounds of formula I in which Y1 and Y2 are each O, X is NR1(R1=H), Z═N(R7)SO2, m=1, R4=H and R2, R3, R5 and R7 are as described informula I may be prepared by reacting a compound of formula VIII inwhich R2, R3, R5, R7 and A are as described in formula I, with sulfonylchlorides of formula IX in polar aprotic solvents such as THF or DMF inthe presence of bases such as alkali carbonates or tertiary alkyl aminesor polymeric amines.

Amines of formula VIII are well known in the literature and areavailable from numerous commercial sources. Specific new variations ofcompounds of formula VIII may be made according to known processes bythose skilled in the art. The sulfonyl chlorides of formula IX may beprepared by chlorine oxidation of sulfides or disulfides of formula X,where R8 is a group such as hydrogen, isopropyl, benzyl or a sulfidesuch that formula X comprises of a symmetrical disulfide.

Sulfides of formula X may be made from cysteine or cystine (R2, R3=H)and their esters by sequential treatment with alkali cyanate and strongacids like potassium cyanate and hydrochloric acid. Alternatively,sulfides of formula X may be prepared by subjecting ketones of formulaXI to conditions as described in the transformation of VII to VIIa abovein (a).

The compounds of the invention may be evaluated for example in thefollowing assays:

Isolated Enzyme Assays

Matrix Metalloproteinase Family Including for Example MMP12, MMP13.

Recombinant human MMP12 catalytic domain may be expressed and purifiedas described by Parkar A. A. et al, (2000), Protein Expression andPurification, 20:152. The purified enzyme can be used to monitorinhibitors of activity as follows: MMP12 (50 ng/ml final concentration)is incubated for 30 minutes at RT in assay buffer (0.1M Tris-HCl, pH 7.3containing 0.1M NaCl, 20 mM CaCl₂, 0.040 mM ZnCl and 0.05% (w/v) Brij35) using the synthetic substrate Mac-Pro-Cha-Gly-Nva-His-Ala-Dpa-NH2 inthe presence or absence of inhibitors. Activity is determined bymeasuring the fluorescence at λex 328 nm and λem 393 nm. Percentinhibition is calculated as follows: % Inhibition is equal to the[Fluorescence_(plus inhibitor)−Fluorescence_(background)] divided by the[Fluorescence_(minus inhibitor)−Fluorescence_(background)].

Recombinant human proMMP13 may be expressed and purified as described byKnauper et al. [V. Knauper et al., (1996) The Biochemical Journal271:1544-1550 (1996)]. The purified enzyme can be used to monitorinhibitors of activity as follows: purified proMMP13 is activated using1 mM amino phenyl mercuric acid (APMA), 20 hours at 21° C.; theactivated MMP13 (11.25 ng per assay) is incubated for 4-5 hours at 35°C. in assay buffer (0.1M Tris-HCl, pH 7.5 containing 0.1M NaCl, 20 mMCaCl₂, 0.02 mM ZnCl and 0.05% (w/v) Brij 35) using the syntheticsubstrate7-methoxycoumarin-4-yl)acetyl.Pro.Leu.Gly.Leu.N-3-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl.Ala.Arg.NH₂in the presence or absence of inhibitors. Activity is determined bymeasuring the fluorescence at λex 328 nm and λem 393 nm. Percentinhibition is calculated as follows: % Inhibition is equal to the[Fluorescence_(plus inhibitor)−Fluorescence_(background)] divided by the[Fluorescence_(minus inhibitor)−Fluorescence_(background)].

A similar protocol can be used for other expressed and purified pro MMPsusing substrates and buffers conditions optimal for the particular MMP,for instance as described in C. Graham Knight et al., (1992) FEBS Lett.296(3):263-266.

Adamalysin Family Including for Example TNF Convertase

The ability of the compounds to inhibit proTNFα convertase enzyme may beassessed using a partially purified, isolated enzyme assay, the enzymebeing obtained from the membranes of THP-1 as described by K. M. Mohleret al., (1994) Nature 370:218-220. The purified enzyme activity andinhibition thereof is determined by incubating the partially purifiedenzyme in the presence or absence of test compounds using the substrate4′,5′-Dimethoxy-fluoresceinylSer.Pro.Leu.Ala.Gln.Ala.Val.Arg.Ser.Ser.Ser.Arg.Cys(4-(3-succinimid-1-yl)-fluorescein)-NH₂in assay buffer (50 mM Tris HCl, pH 7.4 containing 0.1% (w/v) TritonX-100 and 2 mM CaCl₂), at 26° C. for 18 hours. The amount of inhibitionis determined as for MMP13 except λex 490 nm and λem 530 nm were used.The substrate was synthesised as follows. The peptidic part of thesubstrate was assembled on Fmoc-NH-Rink-MBHA-polystyrene resin eithermanually or on an automated peptide synthesiser by standard methodsinvolving the use of Fmoc-amino acids andO-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU) as coupling agent with at least a 4- or 5-fold excess ofFmoc-amino acid and HBTU. Ser¹ and Pro² were double-coupled. Thefollowing side chain protection strategy was employed; Ser¹(But),Gln⁵(Trityl), Arg^(8,12)(Pmc or Pbf), Ser^(9,10,11)(Trityl),Cys¹³(Trityl). Following assembly, the N-terminal Fmoc-protecting groupwas removed by treating the Fmoc-peptidyl-resin with in DMF. Theamino-peptidyl-resin so obtained was acylated by treatment for 1.5-2 hrat 70° C. with 1.5-2 equivalents of4′,5′-dimethoxy-fluorescein-4(5)-carboxylic acid [Khanna & Ullman,(1980) Anal Biochem. 108:156-161) which had been preactivated withdiisopropylcarbodiimide and 1-hydroxybenzotriazole in DMF]. Thedimethoxyfluoresceinyl-peptide was then simultaneously deprotected andcleaved from the resin by treatment with trifluoroacetic acid containing5% each of water and triethylsilane. The dimethoxyfluoresceinyl-peptidewas isolated by evaporation, trituration with diethyl ether andfiltration. The isolated peptide was reacted with4-(N-maleimido)-fluorescein in DMF containing diisopropylethylamine, theproduct purified by RP-HPLC and finally isolated by freeze-drying fromaqueous acetic acid. The product was characterised by MALDI-TOF MS andamino acid analysis.

Natural Substrates

The activity of the compounds of the invention as inhibitors of aggrecandegradation to may be assayed using methods for example based on thedisclosures of E. C. Arner et al., (1998) Osteoarthritis and Cartilage6:214-228; (1999) Journal of Biological Chemistry, 274 (10), 6594-6601and the antibodies described therein. The potency of compounds to act asinhibitors against collagenases can be determined as described by T.Cawston and A. Barrett (1979) Anal. Biochem. 99:340-345.

Inhibition of Metalloproteinase Activity in Cell/Tissue Based Activity

Test as an Agent to Inhibit Membrane Sheddases Such as TNF Convertase

The ability of the compounds of this invention to inhibit the cellularprocessing of TNFα production may be assessed in THP-1 cells using anELISA to detect released TNF essentially as described K. M. Mohler etal., (1994) Nature 370:218-220. In a similar fashion the processing orshedding of other membrane molecules such as those described in N. M.Hooper et al., (1997) Biochem. J. 321:265-279 may be tested usingappropriate cell lines and with suitable antibodies to detect the shedprotein.

Test as an Agent to Inhibit Cell Based Invasion

The ability of the compound of this invention to inhibit the migrationof cells in an invasion assay may be determined as described in A.Albini et al., (1987) Cancer Research 47:3239-3245.

Test as an Agent to Inhibit Whole Blood TNF Sheddase Activity

The ability of the compounds of this invention to inhibit TNFαproduction is assessed in a human whole blood assay where LPS is used tostimulate the release of TNFα. Heparinized (10 Units/ml) human bloodobtained from volunteers is diluted 1:5 with medium(RPMI1640+bicarbonate, penicillin, streptomycin and glutamine) andincubated (160 μl) with 20 μl of test compound (triplicates), in DMSO orappropriate vehicle, for 30 min at 37° C. in a humidified (5% CO₂/95%air) incubator, prior to addition of 20 μl LPS (E. coli. 0111:B4; finalconcentration 10 μg/ml). Each assay includes controls of diluted bloodincubated with medium alone (6 wells/plate) or a known TNFα inhibitor asstandard. The plates are then incubated for 6 hours at 37° C.(humidified incubator), centrifuged (2000 rpm for 10 min; 4° C.), plasmaharvested (50-100 μl) and stored in 96 well plates at −70° C. beforesubsequent analysis for TNFα concentration by ELISA.

Test as an Agent to Inhibit In Vitro Cartilage Degradation

The ability of the compounds of this invention to inhibit thedegradation of the aggrecan or collagen components of cartilage can beassessed essentially as described by K. M. Bottomley et al., (1997)Biochem J. 323:483-488.

Pharmacodynamic Test

To evaluate the clearance properties and bioavailability of thecompounds of this invention an ex vivo pharmacodynamic test is employedwhich utilises the synthetic substrate assays above or alternativelyHPLC or Mass spectrometric analysis. This is a generic test which can beused to estimate the clearance rate of compounds across a range ofspecies. Animals (e.g. rats, marmosets) are dosed iv or po with asoluble formulation of compound (such as 20% w/v DMSO, 60% w/v PEG400)and at subsequent time points (e.g. 5, 15, 30, 60, 120, 240, 480, 720,1220 mins) the blood samples are taken from an appropriate vessel intoIOU heparin. Plasma fractions are obtained following centrifugation andthe plasma proteins precipitated with acetonitrile (80% w/v finalconcentration). After 30 mins at −20° C. the plasma proteins aresedimented by centrifugation and the supernatant fraction is evaporatedto dryness using a Savant speed vac. The sediment is reconstituted inassay buffer and subsequently analysed for compound content using thesynthetic substrate assay. Briefly, a compound concentration-responsecurve is constructed for the compound undergoing evaluation. Serialdilutions of the reconstituted plasma extracts are assessed for activityand the amount of compound present in the original plasma sample iscalculated using the concentration-response curve taking into accountthe total plasma dilution factor.

In Vivo Assessment

Test as an Anti-TNF Agent

The ability of the compounds of this invention as ex vivo TNFαinhibitors is assessed in the rat. Briefly, groups of male WistarAlderley Park (AP) rats (180-210 g) are dosed with compound (6 rats) ordrug vehicle (10 rats) by the appropriate route e.g. peroral (p.o.),intraperitoneal (i.p.), subcutaneous (s.c.). Ninety minutes later ratsare sacrificed using a rising concentration of CO₂ and bled out via theposterior vena cavae into 5 Units of sodium heparin/ml blood. Bloodsamples are immediately placed on ice and centrifuged at 2000 rpm for 10min at 4° C. and the harvested plasmas frozen at −20° C. for subsequentassay of their effect on TNFα production by LPS-stimulated human blood.The rat plasma samples are thawed and 175 μl of each sample are added toa set format pattern in a 96 U well plate. Fifty μl of heparinized humanblood is then added to each well, mixed and the plate is incubated for30 min at 37° C. (humidified incubator). LPS (25 μl; final concentration10 μg/ml) is added to the wells and incubation continued for a further5.5 hours. Control wells are incubated with 25 μl of medium alone.Plates are then centrifuged for 10 min at 2000 rpm and 200 μl of thesupernatants are transferred to a 96 well plate and frozen at −20° C.for subsequent analysis of TNF concentration by ELISA.

Data analysis by dedicated software calculates for each compound/dose:

${{Percent}\mspace{14mu}{inhibition}\mspace{14mu}{of}\mspace{14mu}{TNF}\;\alpha} = \frac{{{Mean}\mspace{14mu}{TNF}\;\alpha\mspace{14mu}({Controls})} - {{Mean}\mspace{14mu}{TNF}\;\alpha\mspace{14mu}({Treated}) \times 100}}{{Mean}\mspace{14mu}{TNF}\;\alpha\mspace{14mu}({Controls})}$Test as an Anti-Arthritic Agent

Activity of a compound as an anti-arthritic is tested in thecollagen-induced arthritis (CIA) as defined by D. E. Trentham et al.,(1977) J. Exp. Med. 146:857. In this model acid soluble native type IIcollagen causes polyarthritis in rats when administered in Freundsincomplete adjuvant. Similar conditions can be used to induce arthritisin mice and primates.

Test as an Anti-Cancer Agent

Activity of a compound as an anti-cancer agent may be assessedessentially as described in I. J. Fidler (1978) Methods in CancerResearch 15:399-439, using for example the B16 cell line (described inB. Hibner et al., Abstract 283 p 75 10th NCI-EORTC Symposium, AmsterdamJun. 16-19 1998).

Test as an Anti-Emphysema Agent

Activity of a compound as an anti-emphysema agent may be assessedessentially as described in Hautamaki et al (1997) Science, 277: 2002.

The invention will now be illustrated but not limited by the followingExamples:

General analytical methods: ¹H-NMR spectra were recorded on either aVarian ^(Unity)Inova 400 MHz or Varian Mercury-VX300 MHz instrument. Thecentral solvent peak of chloroform-d (δ_(H) 7.27 ppm),dimethylsulfoxide-d₆ (δ_(H) 2.50 ppm) or methanol-d₄ (δ_(H) 3.31 ppm)were used as internal references. Low resolution mass spectra wereobtained on a Agilent 1100 LC-MS system equipped with an APCI ionizationchamber.

EXAMPLE 1N-{[(4S)-2,5-dioxoimidazolidinyl]methyl}-4-(4-fluorophenoxy)benzenesulfonamideandN-{[(4S)-2,5-dioxoimidazolidinyl]methyl}[1,1′-biphenyl]-4-sulfonamide

To the stirred solution of N-alfa-BOC-(S)-diaminopropionic acid (100 mg,0.5 mmol) in 2.5 ml water containing 0.04 g (0.55 mmol) of sodiumcarbonate was added the soln. of the sulfonyl chloride (0.5 mmol) in 2.5ml of dioxane. The solution was stirred overnight at room temperature,distributed between ethyl acetate (10 ml) and ca 20% citric acid (10ml), the water phase was three times reextracted with ethyl acetate,organic extract was washed with brine, dried, evaporated and the residuewas treated with 4N HCl in dioxane. The mixture was stirred for 20 min,evaporated and dried in vacuo for 4 hrs at 40 C. Then, the residue wasquenched with 3 ml of water solution of sodium carbonate (0.08 g, 0.85mmol) and 0.9 g (1.1 mmol) of potassium cyanate was added and themixture was stirred for 4 hrs at 100 C. After this period, 1 ml of conc.HCl as added, stirred for 1 hr at the same temperature and then allowedto stand at room temperature overnight. The crystals were filtered,washed with dist. water and dried in vacuo (recrystallised from wt.ethanol if necessary)

N-{[(4S)-2,5-dioxoimidazolidinyl]methyl}-4-(4-fluorophenoxy)benzenesulfonamide

MS: m/z=380.1

N-{[(4S)-2,5-dioxoimidazolidinyl]methyl}[1,1′-biphenyl]-4-sulfonamide

MS: m/z=346.1

1H NMR: (DMSO): 3.00m (1.5H), 3.10m (0.6H), (CH₂), 4.10m (1H, CH), 7.5m(3H), 7.70d (2H), 7.4s (4H).

EXAMPLE 2

Compounds of formula I were prepared wherein Y1 is O, Y2 is O, X is NR1,R1 is H, R2 is H, m is 1, R3 is H, R4 is H, Z is SO₂N(R6), R6 is H,(C1-4)alkyl, methylbenzyl, or methylpyridyl, A is a direct bond, and R5varies.

The syntheses were performed in parallel on 20-well plate manuallyoperated. The amino acid (20 um) was dissolved in 5 ml water containing6.36 mg (60 um) of sodium carbonate. 0.5 ml of the solution was pipettedto each well, followed by 0.5 ml of dioxane solution containing 20 um ofcorresponding sulfonyl chloride. The reaction mixture was shaken for 18hrs at room temperature, diluted with 2 ml of methanol and treated with20 mg of Lewatite S100 in each well (acid form) for 5 min. Then allreaction mixtures was filtered, evaporated in vacuo and the evaporatewas treated with 1 ml of 4 N HCl in dioxane for 30 min, evaporated invacuo and 0.5 ml of 0.5 M wt. solution of potassium cyanate was addedand heated to 100° C. for 3 hrs. Then 10 mg of Lewatite S100 (acid form)was added to each well after being cooled to room temperature, followedby 2 ml of methanol, evaporated in vacuo and threated withtrifluoroacetic acid at 80° C. for 2 hrs. After being evaporated, theresidue was purified by flash chromatography on silica using ethylacetate-methanol gradient (up to 10% MeOH). The purity and mol. weightwas monitored by HPLC-MS. Yields: 0.5-1 mg per each well.

5-(2-Methyl-thiazol-5-yl)-thiophene-2-sulfonic acid(2,5-dioxo-imidazolidin-4-ylmethyl)-amide

LC-MS (APCI) M⁺+H⁺=373.4 (m/z).

3-(4-Chloro-phenoxy)N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=396.8 (m/z).

4-(4-Chloro-phenoxy)N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=396.8 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=392.6 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-3-(4-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=392.6 (m/z).

5-(5-TrifluoromethyH-pyrazol-3-yl)-thiophene-2-sulfonic acid(2,5-dioxo-imidazolidin-4-ylmethyl)-amide

LC-MS (APCI) M⁺+H⁺=410.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyp-4-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=376.4 (m/z).

3-(3,4-Dichloro-phenoxy)-N-(dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=430.6 (m/z).

4-(3,4-Dichloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=430.6 (m/z).

4′-Fluoro-biphenyl-4-sulfonic acid(2,5-dioxo-imidazolidin-4-ylmethyl)-amide

LC-MS (APCI) M⁺+H⁺=364.4 (m/z).

5-Pyridin-2-yl-thiophene-2-sulfonic acid(2,5-dioxo-imidazolidin-4-ylmethyl)-amide

LC-MS (APCI) M⁺+H⁺=353.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(2-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=392.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-3-(2-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=430.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-3-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=430.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=430.4 (m/z).

4′-Trifluoromethyl-biphenyl-4-sulfonic acid(2,5-dioxo-imidazolidin-4-ylmethyl)-amide

LC-MS (APCI) M⁺+H⁺=414.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-o-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=376.4 (m/z).

4-(3,5-Dichloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=431.3 (m/z).

4-(2-Chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=396.8 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-3-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=376.4 (m/z).

4-(4-Cyano-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=387.4 (m/z).

4-(4-Cyano-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=401.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-methyl-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=444.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=458.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-isopropyl-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=472.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-isobutyl-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI), M⁺+H⁺=486.5 (m/z).

N-Benzyl-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-4-(4-trifluoromethyl-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=520.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-pyridin-3-ylmethyl-4-(4-trifluoromethyl-phenoxy)-benzene

LC-MS (APCI) M⁺+H⁺=521.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-fluoro-phenoxy)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=394.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(4-fluoro-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=408.4 (m/z).

N-Benzyl-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-4-(4-fluoro-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=470.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-fluoro-phenoxy)-N-pyridin-3-ylmethyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=471.5 (m/z).

4-(4-Chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=410.5 (m/z).

4-(4-Chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-ethyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=424.88 (m/z).

4-(4-Chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-isopropyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=424.88 (m/z).

N-Benzyl-4-(4-chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=486.9 (m/z).

4-(4-Chloro-phenoxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-pyridin-3-ylmethyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=487.9 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-methyl-4-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=390.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=404.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-isopropyl-4-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=418.5 (m/z).

N-Benzyl-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-4-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=466.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-pyridin-3-ylmethyl-4-p-tolyloxy-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=467.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-methoxy-phenoxy)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=406.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(4-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=420.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-isopropyl-4-(4-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=433.5 (m/z).

N-Benzyl-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-4-(4-methoxy-phenoxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=482.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(4-methoxy-phenoxy)-N-pyridin-3-ylmethyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=483.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=363.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-methyl-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=377.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=363.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=363.5 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(pyridin-2-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=376.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(pyridin-2-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=391.4 (m/z).

4-(5-Chloro-pyridin-2-yloxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=397.8 (m/z).

4-(5-Chloro-pyridin-2-yloxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=410.8 (m/z).

4-(5-Chloro-pyridin-2-yloxy)-N-(2,5-dioxo-imidazolidin-4-ylmethyl)-N-ethyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=425.8 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-N-ethyl-4-(5-fluoro-pyrimidin-2-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=409.8 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(5-fluoro-pyrimidin-2-yloxy)-N-methyl-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=396.4 (m/z).

N-(2,5-Dioxo-imidazolidin-4-ylmethyl)-4-(5-fluoro-pyrimidin-2-yloxy)-benzenesulfonamide

LC-MS (APCI) M⁺+H⁺=382.4 (m/z).

EXAMPLE 3

Compounds were prepared according to Scheme 2 as shown in thedescription above.

(a) Preparation of Starting Materials (Aldehydes or Ketones)

Aldehydes were prepared according to the procedure described by FehrentzJ A and Castro B, Synthesis, 676, (1983). Ketones were preparedaccording to the procedure described by Nahm S and Weinreb S M:Tetrahedron Lett. 22, 3815, (1981).

(b) Preparation of Intermediate Hydantoins

The aldehyde or ketone (5 mmol) was dissolved in 50% water ethanol (10ml) and 0.55 g (10 mmol) of sodium cyanide and 2.7 g (25 mmol) ofammonium carbonate was added and the mixture was heated in the sealedtube to 80° C. for 6 hrs. Then it was cooled, pH was adjusted to 4 andit was evaporated in vacuo. The residue was distributed between water(10 ml) and ethyl acetate and water phase was 3-times re-extracted withethyl acetate, then evaporated and diastereoisomeres were separated bysilica chromatography (grad.TBME-methanol 0-10% MeOH). The followinghydantoins were prepared.

R-1-(2,5-dioxoimidazolidin-4-S-yl)-ethyl carbamic acid tert. butylester

LC-MS (APCI):) M⁺+M⁺=244.4,) M⁺-56 (isobutylene) 188.6,) M⁺-BOC=144.4(main peak).

H-NMR (CDCl₃.ppm): 1.23d (3H), 1.45s (9.1H), 4.36m (1.1H), 5.30bs(1.1H), 10.1bs (1.3H).

R-1-(4-Methyl-2,5-dioxoimidazolin-4-S-yl)ethyl carbamoic acid

LC-MS (APCI):) M⁺+H⁺=258.3,) M⁺-56 (-isobutylene) 202.3,) M⁺-BOC=158.3(main peak).

H-NMR (CDCl₃.ppm): 1.22d (3H), 1.44s (9.2H), 1.58s (3.1H), 3.95m (0.9H),5.5bs (1.5H), 7.9bs (0.8H).

R-1-(4-Methyl-2,5-dioxoimidazolin-4-R-yl)ethyl carbamoic acidtert-butylester

LC-MS (APCI):) M⁺+H⁺=258.3,) M⁺-56 (-isobutylene) 202.3,) M⁺-BOC=158.3(main peak).

H-NMR (CDCl₃.ppm): 1.29d (3H), 1.54s (9.1H), 1.50s (2.95H), 4.25m(1.1H), 5.5bs (1.8H), 7.9bs (0.6H).

R-1-(2,5-dioxo-4-phenylimidazolidin-4-S-yl)-ethyl carbamoic acidtert-butyl ester

LC-MS (APCI):) M⁺+H⁺=320.3) M⁺-56 (-isobutylene) 264.3,) M⁺-BOC=230.3(main peak).

H-NMR (CDCl₃.ppm): 1.31d (3H), 1.35s (9.2H), 4.65m (0.9H), 6.10d(0.94H), 7.25m (3.2H), 7.60d (2.05H).

tert-butyl(2S)-2-[(4R)-2,5-dioxoimidazolidin-4-yl]pyrrolidine-1-carboxylate

LC-MS: M⁺+H⁺=170.0 (M⁺-BOC).

NMR: (CDCl₃.ppm): 1.26s (9H), 1.7-1.9m (3.37H), 2.1-2.2m (0.84H),3.35-3.44 (1.82H), 4.1bs (1.1H).

tert-butyl(2S)-2-[(4S)-2,5-dioxoimidazolidin-4-yl]pyrrolidine-1-carboxylate

LC-MS: M⁺+H⁺=170.0 (M⁺-BOC).

H-NMR: (CDCl₃.ppm): 1.27s (9H), 1.65-2.0m (broad), (4.47H), 3.55m(1.15H), 3.62m (0.55H), 4.4m (0.87H).

tert-butyl(2R)-2-[(4S)-2,5-dioxoimidazolidin-4-yl]pyrrolidine-1-carboxylate

LC-MS: M⁺+H⁺=170.0 (M⁺-BOC).

H-NMR: (CDCl₃.ppm): 1.47s (9H), 1.7-2.2m (broad) 4.30H, 3.6m (1.12H),3.8m (078H, 3.6m (1.1H).

tert-butyl(2R)-2-[(4R)-2,5-dioxoimidazolidin-4-yl]pyrrolidine-1-carboxylate

LC-MS: M⁻+H⁺=170.0 (M⁺-BOC).

H-NMR: (CDCl₃.ppm): 1.47s (9H), 1.7-2.2M (broad) 4.30H, 3.6m (1.12H),3.8m (078H, 3.6m (1.1H).

tert-butyl(2R)-2-[(4S)-4-methyl-2,5-dioxoimidazolidin-4-yl]pyrrolidine-1-carboxylate

LC-MS: M⁺+H⁺=183.1 (M⁺-BOC).

H-NMR: (CDCl₃.ppm): 1.4s (9H) 1.50s (3.2H), 1.65-2.1m (broad) 4.20H,3.4m (1.1H), 3.5bs (0.78H, 4.4m (0.94H).

Deprotection of BOC protected hydantoins was performed via 40%trifluoroacetic acid in DCM and the final compound 5-(1-aminoethyl)5-alkyl imidazoline-2,4 dione trifluoracetate was precipitated by etherafter evaporated to dryness.

R-5-(S-1-aminoethyl)-imidazoline-2,4-dione trifluoroacetate

LC-MS (APCI): M⁺+H⁺=144.2 (m/z).

R-5-(1-aminoethyl)-5-S-methyl imidazolidine-2,4-dione trifluoroacetate

LC-MS (APCI): M⁺+H⁺=158.2 (m/z).

R-5-(1-aminoethyl)-5-R-methyl imidazolidine-2,4-dione trifluoroacetate

LC-MS (APCI): M⁺+H⁺=158.2 (m/z).

R-5-(1-aminoethyl)-5-S-phenylimidazolidine-2,4-dione trifluoroacetate

LC-MS (APCI): M⁺+H⁺=220.3 (m/z).

(5R)-5-[(2S)-pyrrolidin-2-yl]imidazolidine-2,4-dione trifluoroacetate

LC-MS (APCI): M⁺+H⁺=169.1 (m/z).

(5R)-5-[(2R)-pyrrolidin-2-yl]imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=169.1 (m/z).

(5R)-5-[(2S)-pyrrolidin-2-yl]imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=169.1 (m/z).

(5S)-5-[(2S)-pyrrolidin-2-yl]imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=169.1 (m/z).

(5S)-5-methyl-5-[(2R)-pyrrolidin-2-yl]imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=183.21 (m/z).

(c) Preparation of Hydantoins of Formula I

Synthesis was performed in parallel, on 20 well plates, manuallyoperated. Each well was charged by ca 7.5 umol of the correspondingsulfonyl chloride in 0.5 ml of DCM, followed by ca 15-20 umol of the5-(1-aminoethyl) 5-alkyl imidazoline-2,4-dione trifluoroacetate in 0.5ml DCM (small amount of DMF added if necessary for complete dissolution)and 10 mg of the diethylaminomethyl polystyrene resin was added. Themixture was shaked overnight, filtered through 200 mg of silica gel(washed with 3-5 ml of ethyl acetate and the purity was monitored byLC-MS. The solutions were evaporated to dryness to afford all expectedcompounds in sufficient purity.

4-R-(4-chlorophenoxy-N-(1-(2,5-dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=411.1 (m/z).

4-R-(5-chloropyridin-2-oxy)-N-(1-(2,5-dioxoimidazoline-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=412.1 (m/z).

R-N-(1-(2,5-dioxo-imidazolidin-S-4-yl)ethyl)-4-(pyridin-2-yloxy)-benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=378.9 (m/z).

R-N-(1-(2,5-dioxo-imidazolidin-S-4-yl)ethyl)-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=378.9 (m/z).

4-R-(4-cyanophenoxy-N-(1-(2,5dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=401.5 (m/z).

4-R-(4-fluorophenoxy-N-(1-(2,5dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=394.3 (m/z).

4-R-(4-trifluoromethylphenoxy-N-(1-(2,5-dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=444.4 (m/z).

4-R-(4-methylphenoxy-N-(1-(2,5-dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=389.43 (m/z).

4-R-(4-methoxyphenoxy-N-(1-(2,5dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=406.4 (m/z).

4-R-(4-phenoxy-N-(1-(2,5dioxoimidazolin-4-S-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=376.2 (m/z).

R-N-(1-(4-methy2,5-dioxo-imidazolidin-4-S-yl)-ethyl-4-phenoxybenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=390.4 (m/z).

4-(4-Chlorohenoxy-N-(1-(4-S-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=423.4 (m/z).

4-(5-chloropyridyl-2-oxy)-N-(1-(4-S-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=424.4 (m/z).

N-(1-(4-S-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethyl)-4-(pyridin-2-yloxy)benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=392.4 (m/z).

N-(1-(4-S-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethyl)-4-(pyridin-2-yloxy)benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=392.4 (m/z).

4-(4-cyanophenoxy-N-(1-(4-S-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=415.4 (m/z).

R-N-(1-(4-methy2,5-dioxo-imidazolidin-4-R-yl)-ethyl-4-phenoxybenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=390.4 (m/z).

4-(4-Chlorohenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=423.4 (m/z).

4-(5-chloropyridyl-2-oxy)-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=424.4 (m/z).

N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethyl)-4-(pyridin-2-yloxy)benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=392.4 (m/z).

N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethyl)-4-(pyridin-2-yloxy)benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=392.4 (m/z).

4-(4-cyanophenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=415.4 (m/z).

4-(4-fluorophenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-S-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=407.4 (m/z).

4-(4-trifluoromethylphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-S-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=458.4 (m/z).

4-(4-Methylphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-S-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=404.5 (m/z).

4-(4-Methoxyphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-S-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=420.5 (m/z).

4-(4-Phenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-S-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=390.5 (m/z).

4-(4-fluorophenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=407.4 (m/z).

4-(4-trifluoromethylphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=458.4 (m/z).

4-(4-Methylphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=404.5 (m/z).

4-(4-Methoxyphenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=420.5 (m/z).

4-(4-Phenoxy-N-(1-(4-R-methyl-2,5-dioxoimidazolidin-4-R-yl)-ethylbenzenesulfonamide

LC-MS (APCI): M⁺+H⁺=390.5 (m/z).

4-(4-Chlorophenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=486.8 (m/z).

4-(5-chloropyridin-2-yloxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=487.8 (m/z).

N-(1-S-(2,5-dioxo-4-phenylimidazolidin-4-R-yl)-ethyl-4-(pyridin-2-yloxy)-benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=454.6 (m/z).

N-(1-S-(2,5-dioxo-4-phenylimidazolidin-4-R-yl)-ethyl-4-(pyridin-4-yloxy)-benzenesulfonamide

LC-MS (APCI): M⁺+2H⁺=454.6 (m/z).

4-(4-Cyanophenoxy)-N-(1-((2,5-dioxo-1-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=477.6 (m/z).

4-(4-Fluorophenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=470.5 (m/z).

4-(4-Trifluoromethylphenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=519.1 (m/z).

4-(4-Methylphenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=466.4 (m/z).

4-(4-Methoxyphenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=482.4 (m/z).

4-(4-Phenoxy)-N-(1-((2,5-dioxo-4-S-phenyl-imidazolidin-4-R-yl)-ethyl)benzenesulfonamide

LC-MS (APCI): M⁺+H⁺=452.5 (m/z).

5-(1-{[4-(4-chlorophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=450.5 (m/z).

5-(1-{[4-(4-methoxyphenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=446.2 (m/z).

5-(1-{[4-(4-methylphenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=430.1 (m/z).

5-(1-{[4-(4-fluorophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=434.1 (m/z).

(1-{[4-(4-cyanophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=441.1 (m/z).

5-(1-{[4-(4-chlorophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=436.1 (m/z).

5-(1-{[4-(4-fluorophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=420.1 (m/z).

5-(1-{[4-(4-methylphenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=416.1 (m/z).

5-(1-{[4-(4-methoxyphenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=432.1 (m/z).

5-(1-{[4-(4-cyanophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)imidazolidine-2,4-dione

LC-MS (APCI): M⁺+H⁺=427.1 (m/z).

EXAMPLE 4

[(4R)-2,5-dioxoimidazolidinyl]methanesulfonyl chloride,[(4S)-2,5-dioxoimidazolidinyl]methanesulfonyl chloride or[(R)-2,5-Dioxoimidazolidinyl]-methanesulfonyl chloride was reacted withthe appropriate primary or secondary amine to give the compounds listedbelow. All the amines employed are commercially available.

Sulfonyl chloride (0.060 mmoles), amine (0.060 mmoles), triethylamine(0.0084 mL, 0.060 mmoles) in dry tetrahydrofuran (0.70 mL) were stirredat room temperature over night. Polystyrene methylisocyanate (0.025 g,0.030 mmoles) was added and the mixture was shaken over night. The whitesuspension was filtered and the solids were rinsed with tetrahydrofuran(2×1 mL). The filtrates were evaporated, the white solid was suspendedin water (5 mL), collected on a filter, washed with water (2×1 mL),sucked free of water and dried in vacuo at 45° C. over night to affordthe title compounds.

The starting materials were prepared as follows:

5-methyl-5-{[(phenylmethyl)thio]methyl}imidazolidine-2,4-dione

A steel vessel was charged with ethanol and water (315 mL/135 mL).

31.7 g (0.175 mol) of benzylthioacetone, 22.9 g (0.351 mol) of potassiumcyanide and 84.5 g (0.879 mol) of ammonium carbonate was added. Theclosed reaction vessel was kept in an oil bath (bath temperature 90° C.)under vigorous stirring for 3 h.

The reaction vessel was cooled with ice-water (0.5 h), the yellowishslurry was evaporated to dryness and the solid residue partitionedbetween 400 mL water and 700 mL ethylacetate and separated. Thewater-phase was extracted with ethylacetate (300 mL). The combinedorganic phases were washed with saturated brine (150 mL), dried(Na₂SO₄), filtered and evaporated to dryness. If the product did notcrystallize, 300 mL of dichloromethane was added to the oil. Evaporationgave the product as a slightly yellowish powder, 43.8 g (90%).

LC-MS (APCI) m/z 251.1 (MH+).

¹H NMR (DMSO-d₆) δ: 10.74 (1H, s); 8.00 (1H, s); 7.35-7.20 (5H, m); 3.76(2H, s); 2.72, 2.62 (1H each, ABq, J=14.0 Hz); 1.29 (3H, s).

¹³C NMR (DMSO-d₆) δ: 177.30, 156.38, 138.11, 128.74, 128.24, 126.77,62.93, 37.96, 36.39, 23.15.

(5S)-5-methyl-5-{[(phenylmethyl)thio]methyl}imidazolidine-2,4-dione

The title compound was prepared by chiral separation of the racemicmaterial using a 250 mm×50 mm column on a Dynamic Axial CompressionPreparative HPLC system. The to stationary phase used was CHIRALPAK AD,eluent=Methanol, flow=89 mL/min, temp=ambient, UV=220 nm, sampleconc=150 mg/mL, injection volume=20 mL.

Retention time for title compound=6 min.

Analysis of chiral purity was made using a 250 mm×4.6 mm CHIRALPAK-ADcolumn from Daicel, flow=0.5 mL/min, eluent=Ethanol, UV=220 nm,temp=ambient.

Retention time for title compound=9.27 min.

Purity estimated to >99% ee.

LC-MS (APCI) m/z 251.1 (MH+).

[α]_(D)=−30.3° (c=0.01 g/mL, MeOH, T=20° C.).

¹H NMR (DMSO-d₆) δ: 10.74 (1H, s); 8.00 (1H, s); 7.35-7.20 (5H, m); 3.76(2H, s); 2.72, 2.62 (1H each, ABq, J=14.0 Hz); 1.29 (3H, s).

¹³C NMR (DMSO-d₆) δ: 177.30, 156.28, 138.11, 128.74, 128.24, 126.77,62.93, 37.96, 36.39, 23.15.

(5R)-5-methyl-5-{[(phenylmethyl)thio]methyl}imidazolidine-2,4-dione

The title compound was prepared by chiral separation of the racemicmaterial using a 250 mm×50 mm column on a Dynamic Axial CompressionPreparative HPLC system. The stationary phase used was CHIRALPAK AD,eluent=Methanol, flow=89 mL/min, temp=ambient, UV=220 nm, sampleconc=150 mg/mL, injection volume=20 mL.

Retention time for title compound=10 min.

Analysis of chiral purity was made using a 250 mm×4.6 mm CHIRALPAK-ADcolumn from Daicel, flow=0.5 mL/min, eluent=Ethanol, UV=220 nm,temp=ambient.

Retention time for title compound=17.81 min.

Chiral purity estimated to >99% ee.

LC-MS (APCI) m/z 251.0 (MH+).

[α]_(D)=+30.3° (c=0.01 g/mL, MeOH, T=20° C.).

¹H NMR (DMSO-d₆) δ: 10.74 (1H, s); 8.00 (1H, s); 7.35-7.20 (5H, m); 3.76(2H, s); 2.72, 2.62 (1H each, ABq, J=14.0 Hz); 1.29 (3H, s).

¹³C NMR (DMSO-d₆) δ: 177.31, 156.30, 138.11, 128.74, 128.25, 126.77,62.94, 37.97, 36.40, 23.16.

[(4S)-4-methyl-2,5-dioxoimidazolidin-4-yl]methanesulfonyl chloride

(5S)-5-methyl-5-{[(phenylmethyl)thio]methyl}imidazolidine-2,4-dione(42.6 g; 0.17 mol) was dissolved in a mixture of AcOH (450 mL) and H₂O(50 mL). The mixture was immersed in an ice/water bath, Cl₂ (g) wasbubbled through the solution, the flow of gas was adjusted so that thetemperature was kept below +15° C. After 25 min the solution becameyellow-green in colour and a sample was withdrawn for LC/MS and HPLCanalysis. It showed that starting material was consumed. The yellowclear solution was stirred for 30 min and an opaque solution/slurry wasformed.

The solvent was removed on a rotary evaporator using waterbath withtemperature held at +37° C. The yellowish solid was suspended in Toluene(400 mL) and solvent removed on the same rotary evaporator. This wasrepeated once more.

The crude product was then suspended in iso-Hexane (400 mL) and warmedto +40° C. while stirring, the slurry was allowed to cool to roomtemperature before the insoluble product was removed by filtration,washed with iso-Hexane (6×100 mL), and dried under reduced pressure at+50° C. over night. This gave the product as a slightly yellow powder.

Obtained 36.9 g (95%) of the title compound.

Purity by HPLC=99%, NMR supported that purity.

[α]_(D)=−12.4° (c=0.01 g/mL, THF, T=20° C.).

¹H NMR (THF-d₃): δ 9.91 (1H, bs); 7.57 (1H, s); 4.53, 4.44 (1H each,ABq, J=14.6 Hz); 1.52 (s, 3H, CH₃).

¹³C NMR (THF-dg): δ 174.96; 155.86; 70.96; 61.04; 23.66.

[(4R)-4-methyl-2,5-dioxoimidazolidin-4-yl]methanesulfonyl chloride

Following the procedure described for[(4S)-4-methyl-2,5-dioxoimidazolidin-4-yl]methanesulfonyl chloride.

Starting from(5R)-5-methyl-5-{[(phenylmethyl)thio]methyl}imidazolidine-2,4-dione(10.0 g, 40 mmol).

Obtained 8.78 g (96% yield) of the title compound.

Purity by NMR>98%.

[α]_(D)=+ 12.8° (c=0.01 g/mL, THF, T=20° C.).

¹H NMR (THF-d₈): δ 9.91 (1H, brs); 7.57 (1H, s); 4.53, 4.44 (1H each,ABq, J=14.6 Hz); 1.52 (s, 3H, CH₃).

¹³C NMR (THF-dg): δ 174.96; 155.84; 70.97; 61.04; 23.66.

The Table below gives the Amine group for each compound of the abovestructure.

The Table below gives the Amine group for each compound of the abovestructure.

The Table below gives the Amine group for each compound of the abovestructure.

Hydantoin Analysis⁽¹⁾

MW. 375.41 m/z 410 (MH+)

m/z 374 (MH+) MW. 373.43

m/z 388 (MH+) MW. 387.42

N-[4-(4-Chloro-phenoxy)-phenyl]-C-((4S)-4-methyl-2,5-dioxo-imidazolidin-4-yl)-methanesulfonamide

LC-MS (APCI) m/z 410 (MH+).

¹H NMR (DMSO-d₆): δ 10.75 (1H, s); 9.89 (1H, s); 8.04 (1H, s); 7.45-7.39(2H, m); 7.25-7.19 (2H, m); 7.06-6.97 (4H, m); 3.54 (1H from ABq, J=14.1Hz); 1.31 (3H, s).

N-(4-Benzyl-phenyl)-C-((4S)-4-methyl-2,5-dioxo-imidazolidin-4-yl)-methanesulfonamide

LC-MS (APCI) m/z 374 (MH+).

¹H NMR (DMSO-d₆): δ 10.74 (1H, s); 9.82 (1H, s); 8.01 (1H, s); 7.33-7.05(9H, m); 3.49, 3.36 (1H each, ABq, J=16.2 Hz); 1.28 (3H, s).

N-(4-Benzoyl-phenyl)-C-((4S)-4-methyl-2,5-dioxo-imidazolidin-4-yl)-methanesulfonamide

LC-MS (APCI) m/z 388 (MH+).

¹H NMR (DMSO-d₆): δ 10.81 (1H, s); 10.58 (1H, s); 8.08 (1H, s);7.76-7.62 (5H, m); 7.60-7.52 (2H, m); 7.33-7.27 (2H, m); 3.68, 3.52 (1Heach, ABq, J=14.7 Hz); 1.33 (3H, s).

EXAMPLE 5

Prepared from commercially available N-Boc-4-piperidone by methodsdescribed in Example 3.

What we claim is: 1.(1-{[4-(4-cyanophenoxy)phenyl]sulfonyl}pyrrolidin-2-yl)-5-methylimidazolidine-2,4-dione

or a pharmaceutically acceptable salt thereof.
 2. A pharmaceuticalcomposition which comprises a compound or salt as claimed in claim 1 anda pharmaceutically acceptable carrier.